Alikhani Mehdi, Shafaie Ebrahim, Mirabzadeh Ardakani Esmat, Esmaeili Maryam, Saberi Samaneh, Hatefi Mojgan, Mohammadi Marjan
HPGC Research Group, Medical Biotechnology Department, Pasteur Institute of Iran, Tehran, Iran.
Animal Sciences Lab, Department of Molecular Medicine, Pasteur Institute of Iran, Tehran, Iran.
Iran Biomed J. 2019 Jul;23(4):297-302. doi: 10.29252/.23.4.297. Epub 2018 Jul 25.
Quantitation of Helicobacter pylori (Hp) in the gastric tissue is essential for assessment of vaccination/therapeutic regimens.
MATERIALS & RESULTS: Here, the inhibitory effect of mouse gastric DNA (MgDNA) on amplification of Hp genomic DNA (HpDNA) was evaluated by spiking HpDNA with serial dilutions of MgDNA, which yielded concentrations of >10 ng/µl and >0.63-10 ng/µl of MgDNA, as inhibition and interference zones, respectively. Mice were then inoculated with varying doses of Hp and assessed at the inhibition-free concentration of 0.63 ng/µl. The average Hp copy numbers per microgram of gastric tissue discriminated mice having received high vs. low dose inoculums (p < 0.001). Secondly, Hp copy numbers were quantitated in immunized mice, which demonstrated significantly lower numbers, in reference to controls (p = 0.006).
Our method, bypassing the inhibition and/or interference imposed by MgDNA, was able to quantitate gastric tissue-colonizing Hp, segregating mice inoculated with low vs. high doses of Hp, as well as those immunized from controls.
对胃组织中的幽门螺杆菌（Hp）进行定量分析对于评估疫苗接种/治疗方案至关重要。
在此，通过将Hp基因组DNA（HpDNA）与一系列稀释的小鼠胃DNA（MgDNA）混合，评估了MgDNA对HpDNA扩增的抑制作用，分别产生浓度>10 ng/µl和>0.63 - 10 ng/µl的MgDNA作为抑制区和干扰区。然后给小鼠接种不同剂量的Hp，并在0.63 ng/µl的无抑制浓度下进行评估。每微克胃组织中的平均Hp拷贝数区分了接受高剂量与低剂量接种物的小鼠（p < 0.001）。其次，对免疫小鼠的Hp拷贝数进行了定量，与对照组相比，其数量显著降低（p = 0.006）。
我们的方法绕过了MgDNA施加的抑制和/或干扰，能够对胃组织定植的Hp进行定量分析，区分接种低剂量与高剂量Hp的小鼠，以及免疫小鼠与对照小鼠。
